Comparative evaluation of molecular methods for the quantitative measure of torquetenovirus viremia, the new surrogate marker of immune competence.

Authors:
Lisa Macera
Lisa Macera
Pisa University Hospital
Pisa | Italy
Chiara Medici
Chiara Medici
Università di Pisa
Italy
Daniele Focosi
Daniele Focosi
University of Pisa
Italy
Paola Mazzetti
Paola Mazzetti
University of Pisa
Italy
David Navarro
David Navarro
Hospital Clínico Universitario
Spain

J Med Virol 2019 Apr 19. Epub 2019 Apr 19.

Department of Translational Research, University of Pisa, Pisa, Italy.

Background: Torquetenovirus (TTV) viremia is emerging as a promising tool to assess functional immune competence, to predict posttransplant immune-related complications, and eventually to customize immunosuppression.

Methods: In this study, 327 blood samples were tested using two real-time PCR (rtPCR) assays both targeted to the untranslated region of the TTV genome. The first assay was an in-house rtPCR developed by our group, the second one was the recently marketed TTV R-GENE assay.

Results: In the validation study, the TTV R-GENE showed good performances in precision and reproducibility, and sensitivity as low as 12 TTV DNA copies/mL, like previously reported for the in-house rtPCR. The Bland-Altman analysis showed that the mean difference between the two methods was -0.3 log copies/mL. In the comparison study, 69% and 72% of samples were detected positive by rtPCR and TTV R-GENE, respectively (94% concordance, κ = 0.88). Performances did not differ between the two rtPCRs by type of TTV group examined. When a newly-developed in-house digital droplet PCR was applied for TTV quantification and used as an alternative method of comparison on 94 samples, the results strongly correlated with those obtained by the two rtPCR methods (99% concordance).

Conclusion: In summary, all the molecular methods assayed are highly sensitive and accurate in quantitation of TTV DNA in blood samples.

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Source
https://onlinelibrary.wiley.com/doi/abs/10.1002/jmv.25488
Publisher Site
http://dx.doi.org/10.1002/jmv.25488DOI Listing
April 2019
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References

(Supplied by CrossRef)
Classification of TTV and related viruses (anelloviruses)
Biagini P et al.
Curr Top Microbiol Immunol 2009

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